Development 143: doi:10.1242/dev.129122: Supplementary information
Supplementary Figures
Figure S1. Expression of CD68 in mid-embryonic and adult lung macrophages.
A,B, Sections of embryonic (E15.5, A) and adult (P63, B) c-fms-EGFP mouse lungs
immunostained for GFP (green) to show cytoplasm of c-fms-expressing macrophages,
A) or endothelial marker PECAM (grey, B), and counterstained with DAPI (blue, nuclei).
CD68 is expressed in all c-fms-EGFP positive macrophages. CD68 expression at E15.5
is speckled and dim (A), whereas in the adult it is more prominent (B). Bar, 20 um (A),
40 um (B)
Development • Supplementary information
and for macrophage marker microsialin CD68 (red), epithelial marker E-cadherin (grey,
Development 143: doi:10.1242/dev.129122: Supplementary information
Figure S2. Mac2 and MHCII expression in complementary populations of adult
lung macrophages. Sections of adult (P67) c-fms-EGFP mouse lungs immunostained
for EGFP (c-fms, green) to show cytoplasm of c-fms-expressing macrophages, and for
macrophage markers Mac2 (red, A-C) or MHCII (red, D-F), endothelial marker PECAM
(grey), and counterstained with DAPI (blue). B and C are close-ups of boxed regions in
A, and E and F are close ups of boxed regions in D. Note Mac2 is expressed by all of the
alveolar macrophages (A, B) but none of the interstitial macrophages (A, C), and
the alveolar macrophages (E, F). Bar, 40 um (A), 10 um (C), 40 um (D), 10 um (F).
Development • Supplementary information
conversely, MHCII is expressed by all of the interstitial macrophages (D, F) but none of
Development 143: doi:10.1242/dev.129122: Supplementary information
Figure S3. Lineage trace of yolk sac precursors in the juvenile lung. Lineage trace of
yolk sac precursors as in Figure 4 using Runx1-CreER>TdTomato (yolk sac) lineage
trace (Runx1 lineage, red) induced at E6.75, but lung harvested at juvenile stage
(postnatal day P7) and then immunostained for CD68 (green) to visualize all
macrophages, for PECAM (blood vessels, grey), and counterstained with DAPI (blue).
Peripheral (A) and more internal (B) lung sections are shown. Note that as in the adult
lung (Figure 4), yolk sac derived (Runx1 lineage-labeled) macrophages (yellow) localize
surrounding a large blood vessel (B), indicating that the change from broad interstitial to
restricted distribution of yolk sac derived macrophages occurs during the first week of
postnatal life. Bar, 40 um
Development • Supplementary information
to the interstitium peripherally near the mesothelium (meso, A) and more internally
Development 143: doi:10.1242/dev.129122: Supplementary information
Table S1. Summary of marker expression in lung macrophages
csf1r-EGFP
(c-fms,
MacGreen)
Cd68
Lyz2 d
(Lyz)
Emr1
(Adre1,
F4/80)
H2 d
(MHCII)
Lyve1
Lgals3
(Mac2)
Itgax
(CD11c)
Marco
Mrc1
(MMR)
Protein/function
Macrophage colony
stimulating factor 1
receptor; tyrosine kinase
Microsialin;
endosomal/lysosomal
membrane protein
Lysozyme C type M;
secreted bacteriolytic
protein
EGF-like, mucin-like
hormone receptor-like 1;
cell surface glycoprotein
H-2 class II
histocompatibility agent
Lymphatic vessel
endothelial hyaluronic
acid; membrane protein
Galactose-specific lectin
that binds IgE; acute
inflammatory responses
Integrin alpha-X,
receptor for fibrinogen;
cell interactions in
inflammatory responses
Macrophage receptor;
pattern recognition,
bacterial-binding
Macrophage mannose
receptor; endocytosis of
glycoproteins
Alv
M
E13.5
E16.5
Adult
E13.5
E16.5
Adult
+++
+++
+++
+++
+++
+++
+c
++ c
+++ c
+c
++ c
+++ c
+
+/++
+/++
+++
+++
+++
+++ c
++ c
-c
(few)
+e
+/++ e
+/++ e
-
-
+++ c
-
-
-
(many)
+++ c,f
(few)
+++ c,f
-
-
-
-
-c
-c
-c
+++ c
+++ c
+++ c
+
+
+
++
+++
+++
-
-
-
-
(some)
++
(some)
+++
+++
+++
+++, high macrophage (M) marker expression; ++, moderate; +, low; -, none
b
Fraction of macrophages expressing marker: all, no parenthetical note; (many), >60%; (some), 30-60%;
(few), <30%
c
Based on quantitation of >200 csf1r-EGFP+ macrophages
d
Also expressed in alveolar type II cells
e
Expression on Mac2+ Ms variable but increases over time
f
Transient expression in F4/80 macrophages (peaks at E14.5 and largely absent by E16.5)
Abbreviations: M, macrophage; Int, interstitial; Alv, alveolar
a
Development • Supplementary information
M marker
(synonyms)
Marker expression levela, b
Int
F4/80 M
Mac2 M
M